Phosphoramide mustard exposure induces DNA adduct formation and the DNA damage repair response in rat ovarian granulosa cells.
Phosphoramide mustard exposure induces DNA adduct formation and the DNA damage repair response in rat ovarian granulosa cells.
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DOI：
10.1016/j.taap.2014.11.017
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发表时间：
2015-02-01
影响因子：
3.8
通讯作者：
Keating, Aileen F.
中科院分区：
文献类型：
--
作者：
Ganesan, Shanthi;Keating, Aileen F.
磷酰胺芥末(PM)是抗癌剂环磷酰胺(CPA)的卵毒代谢物，通过与DNA形成NOR-G-OH、NOR-G和G-NOR-G加合物，破坏快速分裂的细胞，可能导致DNA损伤。先前的研究表明，PM可引起大鼠卵巢DNA损伤。为探讨PM是否诱导dna加合物的形成、dna损伤和dna修复反应，用1%二甲基亚砜或PM(3或6μM)处理大鼠自发永生化颗粒细胞(SIGCs)24或48 h，3或6μM PM作用48 h后细胞存活率下降(P<0.05)。6μM PM作用24 h后即可检测到Nor-G-OHDNA加合物，而两种浓度PM作用48 h后均可形成细胞毒性更强的G-Nor-GDNA加合物。γ双链断裂的标志物磷酸化H_2AX(DNAH_2AX)也在PM暴露后增加，与DNA加合物的形成相一致。此外，还观察到参与γ修复的基因(ATM、PARP1、PrKDC、XrCC6和BRCA1)和蛋白质(ATM、DARP1HAX、PARP1、PRKDC、XRCC6和BRCA1)的诱导都具有时间和剂量依赖性。这些数据支持PM诱导卵巢颗粒细胞DNA加合物的形成，诱导DNA损伤，并引发卵巢DNA修复反应。
Phosphoramide mustard (PM), the ovotoxic metabolite of the anti-cancer agent cyclophosphamide (CPA), destroys rapidly dividing cells by forming NOR-G-OH, NOR-G and G-NOR-G adducts with DNA, potentially leading to DNA damage. A previous study demonstrated that PM induces ovarian DNA damage in rat ovaries. To investigate whether PM induces DNA adduct formation, DNA damage and induction of the DNA repair response, rat spontaneously immortalized granulosa cells (SIGCs) were treated with vehicle control (1% DMSO) or PM (3 or 6 μM) for 24 or 48 h. Cell viability was reduced (P < 0.05) after 48 h of exposure to 3 or 6 μM PM. The NOR-G-OH DNA adduct was detected after 24 h of 6 μM PM exposure, while the more cytotoxic G-NOR-G DNA adduct was formed after 48 h by exposure to both PM concentrations. Phosphorylated H2AX (γH2AX), a marker of DNA double stranded break occurrence, was also increased by PM exposure, coincident with DNA adduct formation. Additionally, induction of genes (Atm, Parp1, Prkdc, Xrcc6, and Brca1) and proteins (ATM, γH2AX, PARP-1, PRKDC, XRCC6, and BRCA1) involved in DNA repair were observed in both a time- and dose-dependent manner. These data support that PM induces DNA adduct formation in ovarian granulosa cells, induces DNA damage and elicits the ovarian DNA repair response.
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影响因子：
5.3
作者：
Bindra, RS;Schaffer, PJ;Glazer, PM
通讯作者：
Glazer, PM
影响因子：
2
作者：
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通讯作者：
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影响因子：
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作者：
Ganesan, Shanthi;Bhattacharya, Poulomi;Keating, Aileen F.
通讯作者：
Keating, Aileen F.
影响因子：
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作者：
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通讯作者：
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